The Northeast Structural Genomics Consortium (NESG) , established in 2000, has pioneered high throughput (HTP) protein structure determination, demonstrating that HTP protein production is feasible and benefits tremendously from economies of scale. The long term goal of the NESG is to determine representative three-dimensional structures of the repertoire of protein domain families that constitute higher eukaryotic proteomes, with particular emphasis on the human proteome. In order to achieve this goal, the NESG will continue to develop a scalable and efficient platform for HTP protein sample and structure production. NESG proposes an expansion of its current platform, which will allow protein structure production by X-ray crystallography and nuclear magnetic resonance spectroscopy at rates of 150 - 200 protein structures per year, providing some 850 - 900 non-redundant protein structures over a 5 yr period. Technology development will focus on production and structure analysis of eukaryotic proteins using robotic methods. The NESG protein targets are selected from large protein domain sequence families so as to provide representative structures from these families, and to improve our understanding of protein sequence-structure space. Targets will also be prioritized based on functional relationships, using biological network analyses of co-functioning proteins involved in human cancers and developmental diseases. The high scientific value of our structure analysis efforts will be ensured by cutting edge bioinformatics activities, including target selection, protein interaction network analysis, computational structure/function annotation, homology modeling, and advanced data management and mining activities. The many methods and technologies for structural genomics research developed in this project will provide the next-generation tools for traditional hypothesis-driven structural biology research, and will thus have powerful and broad impact on the infrastructure for scientific research in the United States.